Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1995 Jan;69(1):528–533. doi: 10.1128/jvi.69.1.528-533.1995

Human immunodeficiency virus type 1 Nef induces accumulation of CD4 in early endosomes.

O Schwartz 1, A Dautry-Varsat 1, B Goud 1, V Maréchal 1, A Subtil 1, J M Heard 1, O Danos 1
PMCID: PMC188604  PMID: 7983750

Abstract

We have studied the fate of CD4 in CEM T cells expressing a human immunodeficiency virus type 1 HIV-1 Nef protein. Nef triggered a rapid endocytosis and a degradation of CD4, while most of the p56lck was upheld at the cell membrane. In the presence of Nef, CD4 accumulated in acidic intracellular vesicles that were not stained by antibodies against rab6, a marker of the Golgi apparatus complex. Detection of transferrin in CD4-containing vesicles showed that CD4 was trapped in early endosomes, without significant accumulation of CD4 in late endocytic compartments. Internalization pathways taken by CD4 in Nef+ cells may therefore be different from those observed after treatment with phorbol esters.

Full Text

The Full Text of this article is available as a PDF (740.2 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Acres R. B., Conlon P. J., Mochizuki D. Y., Gallis B. Rapid phosphorylation and modulation of the T4 antigen on cloned helper T cells induced by phorbol myristate acetate or antigen. J Biol Chem. 1986 Dec 5;261(34):16210–16214. [PubMed] [Google Scholar]
  2. Aiken C., Konner J., Landau N. R., Lenburg M. E., Trono D. Nef induces CD4 endocytosis: requirement for a critical dileucine motif in the membrane-proximal CD4 cytoplasmic domain. Cell. 1994 Mar 11;76(5):853–864. doi: 10.1016/0092-8674(94)90360-3. [DOI] [PubMed] [Google Scholar]
  3. Anderson R. G., Falck J. R., Goldstein J. L., Brown M. S. Visualization of acidic organelles in intact cells by electron microscopy. Proc Natl Acad Sci U S A. 1984 Aug;81(15):4838–4842. doi: 10.1073/pnas.81.15.4838. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Anderson S. J., Lenburg M., Landau N. R., Garcia J. V. The cytoplasmic domain of CD4 is sufficient for its down-regulation from the cell surface by human immunodeficiency virus type 1 Nef. J Virol. 1994 May;68(5):3092–3101. doi: 10.1128/jvi.68.5.3092-3101.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Benson R. E., Sanfridson A., Ottinger J. S., Doyle C., Cullen B. R. Downregulation of cell-surface CD4 expression by simian immunodeficiency virus Nef prevents viral super infection. J Exp Med. 1993 Jun 1;177(6):1561–1566. doi: 10.1084/jem.177.6.1561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Blum J. S., Fiani M. L., Stahl P. D. Proteolytic cleavage of ricin A chain in endosomal vesicles. Evidence for the action of endosomal proteases at both neutral and acidic pH. J Biol Chem. 1991 Nov 25;266(33):22091–22095. [PubMed] [Google Scholar]
  7. Brady H. J., Pennington D. J., Miles C. G., Dzierzak E. A. CD4 cell surface downregulation in HIV-1 Nef transgenic mice is a consequence of intracellular sequestration. EMBO J. 1993 Dec 15;12(13):4923–4932. doi: 10.1002/j.1460-2075.1993.tb06186.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Crise B., Buonocore L., Rose J. K. CD4 is retained in the endoplasmic reticulum by the human immunodeficiency virus type 1 glycoprotein precursor. J Virol. 1990 Nov;64(11):5585–5593. doi: 10.1128/jvi.64.11.5585-5593.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dautry-Varsat A., Ciechanover A., Lodish H. F. pH and the recycling of transferrin during receptor-mediated endocytosis. Proc Natl Acad Sci U S A. 1983 Apr;80(8):2258–2262. doi: 10.1073/pnas.80.8.2258. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Duprez V., Smoljanovic M., Lieb M., Dautry-Varsat A. Trafficking of interleukin 2 and transferrin in endosomal fractions of T lymphocytes. J Cell Sci. 1994 May;107(Pt 5):1289–1295. doi: 10.1242/jcs.107.5.1289. [DOI] [PubMed] [Google Scholar]
  11. Franchini G., Robert-Guroff M., Ghrayeb J., Chang N. T., Wong-Staal F. Cytoplasmic localization of the HTLV-III 3' orf protein in cultured T cells. Virology. 1986 Dec;155(2):593–599. doi: 10.1016/0042-6822(86)90219-9. [DOI] [PubMed] [Google Scholar]
  12. Garcia J. V., Miller A. D. Serine phosphorylation-independent downregulation of cell-surface CD4 by nef. Nature. 1991 Apr 11;350(6318):508–511. doi: 10.1038/350508a0. [DOI] [PubMed] [Google Scholar]
  13. Goud B., Zahraoui A., Tavitian A., Saraste J. Small GTP-binding protein associated with Golgi cisternae. Nature. 1990 Jun 7;345(6275):553–556. doi: 10.1038/345553a0. [DOI] [PubMed] [Google Scholar]
  14. Griffiths G., Hoflack B., Simons K., Mellman I., Kornfeld S. The mannose 6-phosphate receptor and the biogenesis of lysosomes. Cell. 1988 Feb 12;52(3):329–341. doi: 10.1016/s0092-8674(88)80026-6. [DOI] [PubMed] [Google Scholar]
  15. Guy B., Kieny M. P., Riviere Y., Le Peuch C., Dott K., Girard M., Montagnier L., Lecocq J. P. HIV F/3' orf encodes a phosphorylated GTP-binding protein resembling an oncogene product. Nature. 1987 Nov 19;330(6145):266–269. doi: 10.1038/330266a0. [DOI] [PubMed] [Google Scholar]
  16. Guy B., Rivière Y., Dott K., Regnault A., Kieny M. P. Mutational analysis of the HIV nef protein. Virology. 1990 Jun;176(2):413–425. doi: 10.1016/0042-6822(90)90011-f. [DOI] [PubMed] [Google Scholar]
  17. Haughn L., Gratton S., Caron L., Sékaly R. P., Veillette A., Julius M. Association of tyrosine kinase p56lck with CD4 inhibits the induction of growth through the alpha beta T-cell receptor. Nature. 1992 Jul 23;358(6384):328–331. doi: 10.1038/358328a0. [DOI] [PubMed] [Google Scholar]
  18. Hurley T. R., Luo K., Sefton B. M. Activators of protein kinase C induce dissociation of CD4, but not CD8, from p56lck. Science. 1989 Jul 28;245(4916):407–409. doi: 10.1126/science.2787934. [DOI] [PubMed] [Google Scholar]
  19. Kaminchik J., Bashan N., Itach A., Sarver N., Gorecki M., Panet A. Genetic characterization of human immunodeficiency virus type 1 nef gene products translated in vitro and expressed in mammalian cells. J Virol. 1991 Feb;65(2):583–588. doi: 10.1128/jvi.65.2.583-588.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Marie-Cardine A., Maridonneau-Parini I., Ferrer M., Danielian S., Rothhut B., Fagard R., Dautry-Varsat A., Fischer S. The lymphocyte-specific tyrosine protein kinase p56lck is endocytosed in Jurkat cells stimulated via CD2. J Immunol. 1992 Jun 15;148(12):3879–3884. [PubMed] [Google Scholar]
  21. Pelchen-Matthews A., Armes J. E., Griffiths G., Marsh M. Differential endocytosis of CD4 in lymphocytic and nonlymphocytic cells. J Exp Med. 1991 Mar 1;173(3):575–587. doi: 10.1084/jem.173.3.575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Pelchen-Matthews A., Armes J. E., Marsh M. Internalization and recycling of CD4 transfected into HeLa and NIH3T3 cells. EMBO J. 1989 Dec 1;8(12):3641–3649. doi: 10.1002/j.1460-2075.1989.tb08538.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Pelchen-Matthews A., Boulet I., Littman D. R., Fagard R., Marsh M. The protein tyrosine kinase p56lck inhibits CD4 endocytosis by preventing entry of CD4 into coated pits. J Cell Biol. 1992 Apr;117(2):279–290. doi: 10.1083/jcb.117.2.279. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Pelchen-Matthews A., Parsons I. J., Marsh M. Phorbol ester-induced downregulation of CD4 is a multistep process involving dissociation from p56lck, increased association with clathrin-coated pits, and altered endosomal sorting. J Exp Med. 1993 Oct 1;178(4):1209–1222. doi: 10.1084/jem.178.4.1209. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sanfridson A., Cullen B. R., Doyle C. The simian immunodeficiency virus Nef protein promotes degradation of CD4 in human T cells. J Biol Chem. 1994 Feb 11;269(6):3917–3920. [PubMed] [Google Scholar]
  26. Sattentau Q. J., Dalgleish A. G., Weiss R. A., Beverley P. C. Epitopes of the CD4 antigen and HIV infection. Science. 1986 Nov 28;234(4780):1120–1123. doi: 10.1126/science.2430333. [DOI] [PubMed] [Google Scholar]
  27. Schwartz O., Rivière Y., Heard J. M., Danos O. Reduced cell surface expression of processed human immunodeficiency virus type 1 envelope glycoprotein in the presence of Nef. J Virol. 1993 Jun;67(6):3274–3280. doi: 10.1128/jvi.67.6.3274-3280.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Shin J., Doyle C., Yang Z., Kappes D., Strominger J. L. Structural features of the cytoplasmic region of CD4 required for internalization. EMBO J. 1990 Feb;9(2):425–434. doi: 10.1002/j.1460-2075.1990.tb08127.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Shin J., Dunbrack R. L., Jr, Lee S., Strominger J. L. Phosphorylation-dependent down-modulation of CD4 requires a specific structure within the cytoplasmic domain of CD4. J Biol Chem. 1991 Jun 5;266(16):10658–10665. [PubMed] [Google Scholar]
  30. Skowronski J., Parks D., Mariani R. Altered T cell activation and development in transgenic mice expressing the HIV-1 nef gene. EMBO J. 1993 Feb;12(2):703–713. doi: 10.1002/j.1460-2075.1993.tb05704.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Sleckman B. P., Shin J., Igras V. E., Collins T. L., Strominger J. L., Burakoff S. J. Disruption of the CD4-p56lck complex is required for rapid internalization of CD4. Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7566–7570. doi: 10.1073/pnas.89.16.7566. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Willey R. L., Maldarelli F., Martin M. A., Strebel K. Human immunodeficiency virus type 1 Vpu protein induces rapid degradation of CD4. J Virol. 1992 Dec;66(12):7193–7200. doi: 10.1128/jvi.66.12.7193-7200.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES